Calcium handling in myocardium from amphibian,avian, and mammalian species: the search for two components

Summary Steps involved in excitation-contraction coupling in mammalian myocardium have been derived using a relatively limited number of animal species. However, the use of animal models for investigations into excitation-contraction coupling in normal and disease states has encompassed a wide range of animal species. We addressed the question as to whether excitation-contraction coupling as currently understood applies to intracellular calcium handling in myocardium from multiple mammalian species, amphibian, and avian myocardium. The bioluminescent calcium indicator aequorin was used to record intracellular calcium transients in both ventricular and atrial tissue. We report that in all mammalian and avian species studied the calcium transient recorded in both ventricular and atrial myocardium is monophasic and reflects calcium release and re-uptake by the sarcoplasmic reticulum. In contrast, the Ca²⁺ transient recorded from salamander myocardium is prolonged relative to mammalian and avian myocardium, and appears to reflect in part trans-sarcolemmal calcium entry. Only in diseased myocardium derived from human and swine myocardium was a second component detected in the calcium transient. These data indicate that sarcoplasmic reticulum calcium handling is pivotal in excitation-contraction coupling for multiple species with differing physiologies. Also, in disease states, intracellular calcium handling is often affected with resultant alterations in the time-course and/or configuration of the calcium transient.     

Chicken hypotensive peptide: purification and characterization

We have purified and isolated a novel, hypotensive peptide from avian ventricular tissue. Ventricular homogenates have been shown to exhibit potent hypotensive activity in the avian and mammalian species, with little natriuresis or diuresis. Using avian mean arterial pressure (MAP) as a bioassay, we were able to purify a peptide which decreased MAP 30% in adult, female chickens. Amino acid analysis indicated that it contained 20 amino acids (including two cysteine residues), and was not similar to the amino acid composition of mammalian atrial natriuretic factors, or other known hypotensive peptides.     

Specificity of desmin to avian and mammalian muscle cells.

The extent of invariance and heterogeneity in desmin, the major component of the muscle form of 100 Å filaments, has been investigated in avian and mammalian muscle and nonmuscle cells with two-dimensional gel electrophoresis and indirect immunofluorescence. Desmin from chick, duck and quail, smooth, skeletal and cardiac muscle cells is resolved into two isoelectric variants, α and β, with each possessing the same charge and electrophoretic mobility in all three avian species irrespective of muscle type. Guinea pig and rat muscle desmin resolves into only one variant; it also possesses the same charge and electrophoretic mobility in the two mammalian species, but it is more acidic and slower in electrophoretic mobility than the two avian variants.In immunofluorescence, desmin is localized together with α-actinin along myofibril Z lines. Antibodies to chick smooth muscle desmin, prepared against the protein purified by preparative SDS gel electrophoresis prior to immunization, cross-react with myofibril Z lines in all three avian species. These antibodies do not cross-react with either rat or guinea pig myofibril Z lines. Similarly, they do not cross-react with avian or mammalian nonmuscle cells grown in tissue culture and known to contain cytoplasmic 100 Å filaments.These results demonstrate that desmin is highly conserved within avian muscle cells and within mammalian muscle cells. It is, however, both biochemically and immunologically distinguishable between avian and mammalian muscle cells, and between muscle and nonmuscle cells. We conclude that there are biochemically and immunologically specific forms of desmin for avian and mammalian muscle cells. Furthermore, within a particular vertebrate species, there are at least two separate classes of 100 Å filaments: the muscle class whose major component is desmin, and the nonmuscle class whose major component is distinct from desmin. Taking into consideration the immunological specificity reported by other laboratories for the 100 Å filaments in glial cells, for neurofilaments and for the epidermal 80 Å keratin filaments, we propose that a given vertebrate species contains at least four major distinguishable classes of 100 Å filaments: muscle 100 Å filaments (desmin filaments), glial filaments, neurofilaments and epidermal keratin filaments.     

黑龙江凉水和丰林保护区鸟类和兽类多样性

2013年12月至2015年5月、2014年4月至2015年5月利用红外相机分别对黑龙江凉水保护区和丰林保护区鸟兽多样性进行研究:凉水保护区累计10736个捕获日,获得有效照片总数14726张,兽类独立照片514张,共4目7科11种,鸟类独立照片107张,共4目8科11种;丰林保护区累计7460个捕获日,获得有效照片总数13677张,兽类独立照片638张,共3目6科9种,鸟类独立照片166张,共4目10科16种。凉水保护区实际监测到的鸟兽物种数约为全部鸟兽物种的70%—78%;丰林保护区实际监测到的鸟兽物种数目全部鸟兽物种的80%—89%,保护区内大部分物种被监测到。凉水和丰林保护区兽类相对丰富度最高的均为松鼠(Sciurus vulgaris)和花鼠(Eutamias sibiricus),其次为西伯利亚狍(Capreolus capreolu),鸟类相对丰富度最高的普通鳾(Sitta europaea)和白腹鸫(Turdus pallidus);物种多样性指数显示鸟兽多样性无显著差异;鸟、兽群落相似性指数Cs=0.621,凉水和丰林保护区鸟兽物种有较多重叠。利用红外相机对凉水和丰林保护区鸟兽多样性进行研究发现,兽类中对夜行性小型啮齿类监测不足,监测到的鸟类以林下活动为主。     

Localization of the Novel Neuropolypeptide h3 in Subsets of Tissues from Different Species

Recently we reported the isolation and partial biochemical characterization of a novel polypeptide, h3, from the human brain and liver. Thin-layer isoelectric focusing showed that the polypeptide was ubiquitously distributed throughout the human brain. Immunophosphatase transfer electrophoresis showed that this protein was localized in several mammalian species and different tissues. In addition, h3 or h3-like protein was demonstrated in subsets of tissues from one avian species. Protein h3 was present in epithelial and muscular tissue, as well as in nervous tissue; however, for all species investigated, it was most abundant in CNS and muscle.     

Control enzyme levels in the plasma, brain and liver from wild birds and mammals in Britain

Brain from 47 avian and 17 mammalian species and the liver from 19 avian and 7 mammalian species has been examined for acetyl cholinesterase and nitrophenyl acetate esterase activities. Plasma from 27 avian and 7 mammalian species has been examined for acetyl cholinesterase, cholinesterase, nitrophenyl acetate esterase, glutamate, oxaloacetate transaminase, glutamate pyruvate transaminase, glutamate dehydrogenase, sorbitol dehydrogenase and lactate dehydrogenase activities. The studies have revealed that variations in enzyme activities occur between species but that there are discernible trends within families. The results indicate that comprehensive control enzyme data is necessary in order to assess the effects of exposure to agricultural chemicals in wildlife.     

Cytoplasmic NADP-linked dehydrogenase activities in avian tissues

Cytoplasmic activities of NADP-linked malic enzyme (E.C. 1.1.1.40), glucose-6-phosphate dehydrogenase (E.C. 1.1.1.49) and NADP-linked isocitrate dehydrogenase (E.C. 1.1.1.42) were determined in tissues of selected avian species, and compared with those in mammals. Malic enzyme was generally more active in avian liver and kidney than in the corresponding mammalian tissues. Hepatic activities as high as 200 units/g wet wt and 100 units/g wet wt were recorded in the Nectariniidae and the Ploceidae respectively. Glucose-6-phosphate dehydrogenase was generally less active in avian tissues than malic enzyme. In passerine birds activities were very low indeed, and in most cases spectrophotometrically undetectable. Malic enzyme and glucose-6-phosphate dehydrogenase were highly active in the adipose tissue of mammals but were inactive in the adipose tissue of birds. Marked increases in hepatic malic enzyme and glucose-6-phosphate dehydrogenase activities were associated in birds with premigratory fattening. Activities of isocitrate dehydrogenase were comparable in the corresponding avian and mammalian tissues, including adipose tissue.     

Is the "mammalian" brown fat-specific mitochondrial uncoupling protein present in adipose tissues of birds?

1. Mitochondria were isolated from the furcular, subcutaneous, abdominal, nape and lateral adipose tissue depots of five species of bird (pheasant, Japanese quail, pigeon, house sparrow and great tit) acclimatized to the Northern winter. 2. Mitochondrial proteins were separated by SDS-polyacrylamide gel electrophoresis, blotted onto nitrocellulose membranes, and probed for the presence of the 32,000-33,000 Mr uncoupling protein characteristic of "mammalian" brown adipose tissue, using an anti-(ground squirrel uncoupling protein)serum. 3. Immunoreactivity consistent with uncoupling protein was not detected in mitochondria from any of the avian adipose tissues. Immunoreactivity was, however, evident in mitochondria from perirenal or interscapular adipose tissue from a range of mammals--rats, mice, golden hamsters, Orkney voles, wood mice, pipistrelle bats, wood lemmings, and newborn lambs, cattle, reindeer and red deer. 4. These results provide biochemical evidence that "mammalian-like" thermogenic brown adipose tissue is absent from avian species; adipose tissues in birds appear to be functionally "white".     

Occurrence of avidin-like biotin-binding capacity in various vertebrate tissues and its induction by tissue injury

• 1.1.|Avidin-like biotin-binding capacity was found in the liver and ovary of the common frog and in the kidney or lung of various avian species.
• 2.2. The induction of biotin-binding capacity by tissue injury was marked in various avian species, while in the lizard and in the skin of the common frog it was slight.
• 3.3. Avidin-like biotin-binding capacity or its induction was not found in the tissues of rainbow trout and three mammalian species studied.

     

Effects of cold shock treatment on angiotensin-converting enzyme activity and on semen characteristics in roosters and bulls

Angiotensin-converting enzyme (ACE) activity has been determined in the semen of certain avian and mammalian species as well as its release during cold shock. The maximum and minimum levels of this enzyme were found in mammalian spermatozoa and in seminal plasma, respectively. It was found that ACE activity in mammalian spermatozoa was more pronounced than in the seminal plasma, whereas in the avian species a revers pattern was observed. However, there were no significant differences in ACE activity in spermatozoa and seminal plasma between layer and broiler strains of avian species. By contrast, ACE activity in the spermatozoa and seminal plasma of buffalo bulls was significantly higher (P/ 0.01) than in cattle bulls. Cold shock did not significantly alter semen characteristics in avian species, while a significant (P/ 0.01) decrease in sperm live counts and motility as well as a corresponding increase in morphological abnormalities were observed in the spermatozoa of cattle and buffalo bulls due to cold shock.     

Histochemical comparison of vertebrate mast cells

Synopsis Mast cells from representative species of each of the five vertebrate classes have been studied histochemically. Histamine was found in reptilian, avian and mammalian species. An N-sulfated mucopolysaccharide was present in all species but the carp. A chymotrypsin-like or a trypsin-like esterase or both was found in the turtle and the several mammalian species examined. The mast cell thus appears to have developed the biological armamentarium associated with the mammalian cells in discrete steps during vertebrate evolution.     

Calbindin 28 kDa in endocrine cells of known or putative calcium-regulating function

Summary The distribution of calbindin in some endocrine glands (thyroid, parathyroid, ultimobranchial body, pituitary and adrenals) and in the diffuse endocrine cells of the gut and pancreas has been investigated immunohistochemically using an antiserum raised against the 28 kDa calbindin from chicken duodenum. The identity of calbindin-immunoreactive cells in a number of avian and mammalian species was ascertained by comparison with hormone-reactive cells in consecutive sections or by double immunostaining of the same section with both calbindin and hormone antibodies. Calcitonin-producing C cells of the mammalian and avian thyroid, parathyroid or ultimobranchial body, PP, glucagon and insulin cells of the mammalian and avian pancreas, enteroglucagon cells of the avian intestine, secretin cells of the mammalian duodenum, histamine-producing ECL cells of the mammalian stomach, as well as noradrenaline-producing cells of the adrenal medulla and some (TSH?) cells of the adenohypophysis were among the calbindin-immunoreactive cells. Although some species variability has been observed in the intensity and distribution of the immunoreactivity, especially in the pancreas and the gut, a role for calbindin in the mechanisms of calcium-mediated endocrine cell stimulation or of intracellular and extracellular calcium homeostasis is suggested.     

Calbindin 28 kDa in endocrine cells of known or putative calcium-regulating function. Thyro-parathyroid C cells, gastric ECL cells, intestinal secretin and enteroglucagon cells, pancreatic glucagon, insulin and PP cells, adrenal medullary NA cells and some pituitary (TSH?) cells

The distribution of calbindin in some endocrine glands (thyroid, parathyroid, ultimobranchial body, pituitary and adrenals) and in the diffuse endocrine cells of the gut and pancreas has been investigated immunohistochemically using an antiserum raised against the 28 kDa calbindin from chicken duodenum. The identity of calbindin-immunoreactive cells in a number of avian and mammalian species was ascertained by comparison with hormone-reactive cells in consecutive sections or by double immunostaining of the same section with both calbindin and hormone antibodies. Calcitonin-producing C cells of the mammalian and avian thyroid, parathyroid or ultimobranchial body, PP, glucagon and insulin cells of the mammalian and avian pancreas, enteroglucagon cells of the avian intestine, secretin cells of the mammalian duodenum, histamine-producing ECL cells of the mammalian stomach, as well as noradrenaline-producing cells of the adrenal medulla and some (TSH?) cells of the adenohypophysis were among the calbindin-immunoreactive cells. Although some species variability has been observed in the intensity and distribution of the immunoreactivity, especially in the pancreas and the gut, a role for calbindin in the mechanisms of calcium-mediated endocrine cell stimulation or of intracellular and extracellular calcium homeostasis is suggested.     

Evidence of a true pharyngeal tonsil in birds: a novel lymphoid organ in Dromaius novaehollandiae and Struthio camelus (Palaeognathae)

ABSTRACT: BACKGROUND: Tonsils are secondary lymphoid organs located in the naso- and oropharynx of most mammalian species. Most tonsils are characterised by crypts surrounded by dense lymphoid tissue. However, tonsils without crypts have also been recognised. Gut-associated lymphoid tissue (GALT), although not well-organised and lacking tonsillar crypts, is abundant in the avian oropharynx and has been referred to as the "pharyngeal tonsil". In this context the pharyngeal folds present in the oropharynx of ratites have erroneously been named the pharyngeal tonsils. This study distinguishes between the different types and arrangements of lymphoid tissue in the pharyngeal region of D. novaehollandiae and S. camelus and demonstrates that both species possess a true pharyngeal tonsil which fits the classical definition of tonsils in mammals. RESULTS: The pharyngeal tonsil (Tonsilla pharyngea) of D. novaehollandiae was located on the dorsal free surface of the pharyngeal folds and covered by a small caudo-lateral extension of the folds whereas in S. camelus the tonsil was similarly located on the dorsal surface of the pharyngeal folds but was positioned retropharyngeally and encapsulated by loose connective tissue. The pharyngeal tonsil in both species was composed of lymph nodules, inter-nodular lymphoid tissue, mucus glands, crypts and intervening connective tissue septa. In S. camelus a shallow tonsillar sinus was present. Aggregated lymph nodules and inter-nodular lymphoid tissue was associated with the mucus glands on the ventral surface of the pharyngeal folds in both species and represented the Lymphonoduli pharyngeales. Similar lymphoid tissue, but more densely packed and situated directly below the epithelium, was present on the dorsal, free surface of the pharyngeal folds and represented a small, non-follicular tonsil. CONCLUSIONS: The follicular pharyngeal tonsils in D. novaehollandiae and S. camelus are distinct from the pharyngeal folds in these species and perfectly fit the classical mammalian definition of pharyngeal tonsils. The presence of a true pharyngeal tonsil differentiates these two ratite species from other known avian species where similar structures have not been described. The pharyngeal tonsils in these ratites may pose a suitable and easily accessible site for immune response surveillance as indicated by swelling and inflammation of the tonsillar tissue and pharyngeal folds. This would be facilitated by the fact that the heads of these commercially slaughtered ratites are discarded, thus sampling at these sites would not result in financial losses.     

Host specificity of Cryptosporidium sp. isolated from chickens

The host specificity of Cryptosporidium sp. infecting chickens was evaluated by oral inoculation of oocysts into 6 different species of neonatal rodents, adult nude mice (athymic), neonatal conventional and gnotobiotic pigs, turkeys, muscovy ducks and bobwhite quail. Examinations of tissue sections, ileal mucosal smears, fecal flotations and stained feces failed to reveal any infections in the mammalian species examined. Oocysts were observed in the feces, and developmental stages were observed in tissue sections, of turkeys and muscovy ducks but not bobwhite quail. This study indicates that Cryptosporidium sp. infections in avian species are probably not a zoonotic threat to humans.     

Inhibition of amylases from different origins by albumins from the wheat kernel.

The amylase activity of water extracts from 18 insect species, from 23 marine species and from 17 different species of birds and mammals was determined quantitatively. The inhibition of amylase in these extracts by three albumin fractions from the mature wheat kernel, which had been separated according to their molecular weights (60 000, 24 000 and 12 500 D), was determined as well. The inhibition activity of the three albumin fractions toward amylases extracted from a number of cereal species or from immature and germinating wheat kernel was also tested. The extracts from insects that are destructive of wheat grain and stored wheat products showed much higher amylase activities as compared to the other insect species that do not attack wheat and wheat products. On the basis of the effectiveness with which the three albumin fractions inhibit their activities, the amylase preparations tested were divided into susceptible, partially susceptible and resistent. Susceptible amylases, inhibited by any of the three albumin fractions, were found mainly in insects that attack wheat and in marine species. Partially susceptible amylases, inhibited by only one or two of the three albumin fractions, were present in a few avain and mammalian species including man. Resistent amylases were largely distributed in cereal, avian and mammalian species as well as in insect species that do not usually attack wheat grain or wheat flour products. At no stage of development, wheat alpha-amylase was inhibited by the albumin fractions from the mature kernel. The 12 500 dalton albumin fraction was the most effective in inhibiting insect amylases, but it was inactive toward avian and mammalian amylases. The 24 000 dalton albumin fraction was the most effective in inhibiting amylases from marine avian and mammalian species and inhibited as much as 33 amylases over 66 different amylases tested. It is suggested that protein inhibitors of amylase contributed to natural selection of polyploid wheats by giving some insect resistence to such wheats, even though some insect species were able to overcome this biochemical defense toa large degree by producing higher amylase activities.     

Expression of the Cytoskeletal-Associated Protein Filamin in Adult Rat Organs

Filamin is a well-characterized actin-associated protein first isolated from chicken smooth muscle. Subsequently, this polypeptide and its nonmuscle homolog actin-binding protein have been shown to be expressed in avian muscle tissue, mammalian smooth muscle, mammalian macrophages and other blood cell types, as well as several cultured cell lines. In this report, the occurrence of this polypeptide in adult mammalian organs has been investigated. Immunoblot analysis using three anti-filamin monoclonal antibodies showed that this protein was largely detected in adult rat organs that possess a substantial smooth muscle component. Furthermore, the limited expression of filamin in smooth muscle tissue was corroborated by immunohistochemical analysis. In contrast to avian systems, filamin was never found in detectable quantities in either mammalian cardiac or skeletal muscle. Quantitative immunoblot analysis demonstrated that filamin amounts roughly correlated with the abundance of the smooth muscle component of a given organ, comprising as much as 16.5% of the total SDS-extractable protein in bovine aorta. Work in avian systems and cells in culture has suggested that filamin is a rather ubiquitous cytoskeletal element. By contrast, this work demonstrates that filamin is highly restricted in its expression in mammalian organ systems, in situ.     

Molecular Cloning and Comparison of Avian Preproghrelin Genes

We report cDNA sequences for the preproghrelin gene from goose, duck, and emu. This gene is involved in stimulating the release of growth hormone in mammals and may play a similar role in avian species. The complete coding sequence of avian preproghrelin encodes a 116 amino acid (aa) protein, which is organized into three parts: the N-terminal hydrophobic signal peptide, a 26 aa peptide for mature ghrelin, and a long C-terminal polypeptide. Domain/motif structures of preproghrelin protein are highly conserved among avian species. Although the avian and mammalian homologs are not highly similar for the whole 116 aa sequence, the identity of the highly conserved “active core” sequence and the n-octanoyl modification of the serine 3 residue avian ghrelin protein with its mammalian homologs implies conserved function of ghrelin protein during evolution. Information provided in this study will be useful in further studies to determine the role the preproghrelin gene plays in the regulation of growth hormone release and body weight gain in avian species. Jing Yuan and Jianjun Zhou contributed equally to this work     

Comparative cytomorphology of the avian adrenocortical tissue.

The histology and histochemistry of the interrenal gland of twenty avian species have been described in this report. The avian interrenal tissue, on the basis of nuclear orientation and other cytomorphic features, can be classified into six cytologically distinct types. Cytomorphic organization within the interrenal tissue of these avian species also indicates some regional demarcation into subcapsular and central zones. Zonal demarcation in the interrenal tissue of these birds has also been indicated from chemocytological investigation. Paradoxically not much of correspondence was noticed in the cytologic and cytochemical patterns. The cytological and cytochemical studies fail to indicate any phylogenetic trend within the interrenal tissue of the birds under investigation.     

Efficacy of vaporized hydrogen peroxide against exotic animal viruses.

The efficacy of vapor-phase hydrogen peroxide in a pass-through box for the decontamination of equipment and inanimate materials potentially contaminated with exotic animal viruses was evaluated. Tests were conducted with a variety of viral agents, which included representatives of several virus families (Orthomyxoviridae, Reoviridae, Flaviviridae, Paramyxoviridae, Herpesviridae, Picornaviridae, Caliciviridae, and Rhabdoviridae) from both avian and mammalian species, with particular emphasis on animal viruses exotic to Canada. The effects of the gas on a variety of laboratory equipment were also studied. Virus suspensions in cell culture media, egg fluid, or blood were dried onto glass and stainless steel. Virus viability was assessed after exposure to vaporphase hydrogen peroxide for 30 min. For all viruses tested and under all conditions (except one), the decontamination process reduced the virus titer to 0 embryo-lethal doses for the avian viruses (avian influenza and Newcastle disease viruses) or less than 10 tissue culture infective doses for the mammalian viruses (African swine fever, bluetongue, hog cholera, pseudorabies, swine vesicular disease, vesicular exanthema, and vesicular stomatitis viruses). The laboratory equipment exposed to the gas appeared to suffer no adverse effects. Vaporphase hydrogen peroxide decontamination can be recommended as a safe and efficacious way of removing potentially virus-contaminated objects from biocontainment level III laboratories in which exotic animal disease virus agents are handled.